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B Cell Protocols pp 189–212Cite as

B-Cell Signal Transduction

Tyrosine Phosphorylation, Kinase Activity, and Calcium Mobilization

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 271))

Abstract

Signal transduction by the B-cell antigen receptor (BCR) regulates development, survival, and clonal expansion of B cells. The BCR complex comprises the membrane-bound immunoglobulin molecule and the Ig-α/Ig-β heterodimer, and was shown to form oligomeric structures. Antigen-mediated engagement of the BCR results in the tyrosine phosphorylation of multiple signaling proteins leading to calcium mobilization and the activation of downstream serine/threonine kinases as well as transcription factors. In pervanadate (PV)-treated B cells, comparable pathways are activated on expression of the BCR, indicating that the BCR can signal in an antigen-independent fashion as well. In this chapter, we describe the analysis of antigen-dependent and -independent tyrosine phosphorylation events as well as a method to study calcium mobilization from differentially stimulated B cells. Furthermore, we emphasize the use of phospho-specific antibodies (Abs) and low-molecular-weight enzyme inhibitors in the process of mapping BCR-activated signaling pathways as well as determining activation states of signaling proteins.

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Author information

Authors and Affiliations

  1. Department of Molecular Immunology, Biology III, University of Freiburg and Max-Planck-Institute for Immunobiology, Freiburg, Germany

    Tilman Brummer, Winfried Elis, Michael Reth & Michael Huber

Authors
  1. Tilman Brummer
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  2. Winfried Elis
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  3. Michael Reth
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  4. Michael Huber
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Editor information

Editors and Affiliations

  1. Department of Microbiology, Columbia University College of Physicians and Surgeons, New York, NY

    Hua Gu PhD

  2. The Center for Blood Research, The Harvard Medical School, Boston, MA

    Klaus Rajewsky MD

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© 2004 Humana Press Inc.

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Brummer, T., Elis, W., Reth, M., Huber, M. (2004). B-Cell Signal Transduction. In: Gu, H., Rajewsky, K. (eds) B Cell Protocols. Methods in Molecular Biology, vol 271. Humana Press. https://doi.org/10.1385/1-59259-796-3:189

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  • DOI: https://doi.org/10.1385/1-59259-796-3:189

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-347-3

  • Online ISBN: 978-1-59259-796-3

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